Strategic internal covalent cross-linking of TNF produces a stable TNF trimer with improved TNFR2 signaling

Liqin Ban; Willem Kuhtreiber; John Butterworth; Yoshiaki Okubo; Éva Vanamee; Denise Faustman

Liqin Ban
Willem Kuhtreiber
John Butterworth
Yoshiaki Okubo
Éva Vanamee
Denise Faustman

Keywords: TNF, Trimer, Cell signaling, TNFR2, Transmembrane TNF, Tumor necrosis factor, Tumor necrosis factor receptor

Abstract

Background

Soluble TNF superfamily (TNFSF) ligands are less stable and less active than their transmembrane (tm) analogues. This is a problem for the therapeutic use of recombinant TNFSF ligands in diverse diseases including cancer and autoimmunity. Creating TNFSF ligand analogues with improved targeting of their respective receptors is important for research and therapeutic purposes.

Findings

Covalent internal cross-linking of TNF monomers by double mutations, S95C/G148C, results in stable trimers with improved TNFR2 function. The resulting mutein induced the selective death of autoreactive CD8 T cells in type-1 diabetic patients and demonstrates targeted proliferation and expansion of human CD4 Tregs.

Conclusions

Stable TNF trimers, created by internal covalent cross-linking, show improved signaling. The high structural homology within the TNF superfamily provides an opportunity to extend internal cross-linking to other TNF superfamily proteins to produce active trimers with improved stability and receptor signaling, and with potential applications for cancer, autoimmunity, infections, and transplantation.

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